FR2746641A1 - USE OF AN EXTRACT OF AT LEAST ONE IRIDACEA IN THE TREATMENT OF WRINKLES - Google Patents

Abstract

The present invention relates to the use of an effective amount of at least one extract of at least one lridacea in a cosmetic composition or for the preparation of a pharmaceutical composition, for relaxing and / or relaxing the skin tissue and / or subcutaneous, in particular for treating wrinkles and fine lines of the skin.

Description

The present invention relates to the use of an effective amount

  of at least one extract of at least one Iridaceae in a cosmetic composition or for the preparation of a pharmaceutical composition, for relaxing and / or relaxing the skin and / or subcutaneous tissue, in particular with a view to treating wrinkles and fine lines on the skin. Women, even men, currently tend to want to look young for as long as possible and therefore seek to fade the marks of skin aging, which are reflected in particular by wrinkles and fine lines. In this regard, advertising and fashion refer to products intended to keep glowing, wrinkle-free skin as long as possible, marks of young skin, all the more as the physical appearance induces on the psyche and / or on morale. However, it is important to feel physically and

morally young.

  Until now, wrinkles and fine lines have been treated with cosmetic products containing active agents acting on the skin, for example by moisturizing it or improving its cell renewal or by promoting the synthesis of collagen which makes up the skin tissue. But, to date, we do not know how to act on

  wrinkles by acting on the muscular elements present in the skin.

  It is known that the facial skin muscles are under the control of the motor nerve afferents of the facial nerve and that, moreover, the interlobular partitions of the hypodermis contain within them fibers which constitute striated muscle tissue (panniculus carnosus). On the other hand, it is also known that a subpopulation of fibroblasts of the dermis, which are called myofibroblasts, has characteristics in common with muscle tissue. The Applicant has in particular observed, in certain pathological and therapeutic situations, the role played on facial wrinkles by the nerves controlling all of this muscle tissue. Thus, in attacks of the facial nerve, in which the transmission of nerve impulses is interrupted and / or reduced,

  witness in the innervation territory a paralysis of the facial muscles.

  This facial paralysis is reflected, among other clinical signs, by a

  attenuation or even disappearance of wrinkles.

  Conversely, in states of muscular hyper-contraction of the face, the applicant has observed an accentuation of facial wrinkles. In addition, she also observed an accentuation of facial wrinkles in the states of muscular hypertonia of Parkinson's disease and the side effects induced by neuroleptics. Furthermore, it has been shown that the botulinum toxin, originally used to treat spasms, can act on states of muscle spasticity (see A. Blitzer et al., Arch. Otolaryngol. Head Neck Surg., 1993, 119, pages 1018 to 1022) and on the glabella lines which are the inter-eyebrow lines (see JD Carruters et al., J. Dermatol. Surg. Oncol., 1992, 18, pages 17 to 21). As a result, it is possible

  to act by a pharmacological action on the nervous component of wrinkles.

  In the peripheral nervous system, the junction between a nerve and a muscle constitutes the neuromuscular plate, upstream of which is the afferent nerve path called the motor neuron. Furthermore, the cell membranes of each nerve fiber comprise numerous ion channels, and in particular chlorine channels, capable of letting the corresponding element pass through in ionic form, and in the case of chlorine channels in the form of chloride. These channels are associated with neural receptors. The neural receptors associated with the chlorine channels in the periphery are in particular receptors for glycine (sensitive glycine-strychnine receptors) and receptors for

  GABA type A (GABA-A receptors) (GABA = y-aminobutiric acid).

  It is known that it is possible to decrease the excitability of the motor neuron by various pharmacological agents acting on the sensitive glycinestrychnine receptors or on the GABA-A receptors of the peripheral nervous system (see W. Sieghart,

  Trends in Pharmacological Science, December 1992, vol. 131, pages 446 to 450).

  Thus, the excitability of the motor neuron can be modulated for example by glycine or

  Gamma amino-butyric acid (GABA).

  Activation of these receptors opens the chlorine channels and leads to the entry of chloride ions, which leads to an increase in chloride ions in the nerve fiber cells and therefore to hyperpolarization of the motor neurons which consequently become less excitable. This decrease in motor neuron excitability results in less stimulation of the fiber

  muscle, causing it to relax.

  After numerous clinical tests, the applicant has been able to determine that the contractile muscle fibers, which are under the direct control of the neuro-motor influx, play an essential role in the pathogenesis of wrinkles and that the modulation of the neuro-impulse -motor attenuated not only wrinkles but also fine lines and also had a "smoothing" effect on the skin microrelief. She also found that the skin and subcutaneous tissues contain receptors associated with the chlorine channels, which, until now, had not been envisaged. So she found that we could act on these channels to relax or

  relax these tissues, and thus reduce wrinkles and fine lines.

  No one had, to date, established a link between the chlorine channels of nerve fibers and wrinkles, and had not found that one could treat wrinkles by acting on the chlorine channels by activation of the receptors found in or near these channels. The substances which can activate the receptors of the chlorine channels and thus involve the entry of chloride in the cells, are

called agonist substances.

  There are several receptors associated with the chlorine channel. These include sensitive glycine-strychnine receptors and GABA-A receptors, the latter themselves comprising several subunits consisting of the GABA site, the benzodiazepine site, a type of steroid site and the barbiturate site. All substances acting as agonists of these receptors or sites can be used to relax or relax the skin and / or subcutaneous tissue

according to the invention.

  For a substance to be recognized as a chlorine channel receptor agonist, it must meet the following two characteristics: being able to selectively bind to at least one of the different receptors associated with the chlorine channel;

  - show a relaxation effect on contracted muscle tissue.

  The first characteristic, which consists of the possibility of attaching to a receptor associated with a chlorine channel, does not make it possible to distinguish an agonist activity from an antagonist activity, but it makes it possible to define an affinity

potential for the receiver.

  The second characteristic makes it possible to select the agonists. The agonist activity of the substance studied can be demonstrated by the relaxation effect it produces on muscle tissue which has been previously contracted by a chlorine channel antagonist substance. As a chlorine channel antagonist, it is possible to choose the substances known as such, and

  including the following substances: bicuculline, strychnine, terbutylbicyclo-

  phosphorothionate and picrotoxin. Surprisingly the Applicant has shown that an extract of at least one

  Iridacea met the criteria for agonist of the chlorine channel receptors

defined above.

  Also, the present invention relates to the use, in a cosmetic composition or for the preparation of a pharmaceutical composition, of an effective amount of an extract of at least one Iridaceae to relax and / or relax the

skin and / or subcutaneous tissue.

  A more particular subject of the invention is the use of an effective amount of an extract of at least one Iridaceae to relax and / or relax the skin tissue

  and / or subcutaneous in order to reduce wrinkles and / or fine lines.

  The composition containing the extract according to the invention can be applied either by local route, that is to say by topical route, or by subcutaneous and / or intradermal injection, or by systemic or general route, it is i.e. by mouth

  and / or by intramuscular injection.

  The subject of the invention is also a method of cosmetic treatment of wrinkles and / or fine lines, consisting in applying topically, injecting or ingesting a composition comprising an effective amount of an extract of at least one Iridaceae. The extract of at least one Iridacee can be any extract prepared from material

  plant from the Iridaceae family.

  The composition may contain an extract of at least one Iridaceae obtained from

  plant material from whole plant grown in vivo or from in vitro culture.

  The selection pressure imposed by the physicochemical conditions during the growth of plant cells in vitro makes it possible to obtain a standardized plant material and available throughout the year unlike plants

grown in vivo.

  The term “in vitro culture” means all of the techniques known to those skilled in the art which artificially allow a plant or a part to be obtained.

of a plant.

  It is thus possible, for example, according to the invention, to use an extract of the roots of at least one Iridaceae cultivated in vitro or else an extract of cells.

  undifferentiated by at least one Iridacee.

  Preferably, an extract obtained from plant material cultivated in vitro is used and even more preferably an extract obtained from cells.

  undifferentiated cultivated in vitro.

  By undifferentiated plant cells is meant any plant cell having none of the characteristics of a particular specialization and capable of living by itself and not in dependence on other cells. These undifferentiated plant cells are possibly able, under the effect of an induction, to

  any differentiation consistent with their genome.

  According to the chosen culture method, and in particular according to the chosen culture medium, it is possible to obtain, from the same explant, plant cells

  undifferentiated with different characters.

  The Iridaceae (or Iridae) family has around 750 species.

  Plants of the Iridaceae family are mainly used for their properties

aromatic and ornamental.

  Among the genera of Irridaceae which can be used according to the invention, there may be mentioned by way of example the genera Romulea, Crocus, Iris, Gladiolus, Sisyrinchiumrn or else

Hermodactylus.

  As plant material which can be used, mention may be made of that originating from iris germanica, iris florentina, iris pallida, Crocus versicolor, Romulea bulbucodium or

still from Gladiolus communis.

  More particularly, according to the invention, plant material from the genus is used

  Iris and preferably plant material of iris pallida.

  Any extraction method known to those skilled in the art can be used to

  prepare the Irridaceous extract contained in the composition according to the invention.

  Mention may, in particular, be made of alcoholic extracts, especially ethanolic or

still hydroalcoholic.

  You can also use an Irridaceous extract prepared by the method described

  in French patent application No. 95-02379 filed by the applicant.

  Thus, in a first step the plant material is ground in a cold aqueous solution, in a second step the particles in suspension are removed from the aqueous solution obtained from the first step, and in a third step the aqueous solution obtained from the second step. This

  aqueous solution corresponds to the extract.

  On the other hand, the first step can advantageously be replaced by a simple freezing operation of the plant tissues (for example at 20 ° C.), followed by an aqueous extraction repeating the second and third steps described above. An example of preparation of Irridaceous extract usable according to the invention is

  given elsewhere in the examples.

  The effective amount of Irridaceous extract contained in the composition of the invention is of course a function of the desired effect and can therefore vary

to a large extent.

  To give an order of magnitude, if the composition is a cosmetic composition, it may contain an extract of at least one Iridaceae in an amount representing from 0.01% to 20% of the total weight of the composition and preferably in an amount representing from 0.1% to 5% of the total weight of the composition. To give an order of magnitude, if the composition is a pharmaceutical composition, it may contain an extract of at least one Iridaceae in an amount representing from 0.01% to 30% of the total weight of the composition and preferably in an amount representing 0.1% to 10% of the total weight of

the composition.

  The compositions according to the invention can be presented in all the galenical forms normally used for a topical, injectable or oral application. The amounts of the various constituents of the compositions according to the invention are those conventionally used in the fields under consideration and are appropriate

to their dosage form.

  For topical application, the compositions of the invention comprise a medium compatible with the skin. These compositions can be in particular in the form of aqueous, alcoholic or hydroalcoholic solutions, gels, water-in-oil or oil-in-water emulsions having the appearance of a cream or a gel, of microemulsions, aerosols, or in the form of vesicular dispersions containing ionic and / or nonionic lipids. These dosage forms are prepared according to the usual methods of the fields considered. These compositions for topical application may especially constitute a cosmetic, pharmaceutical or protective, treatment or care composition for the face, for the neck, for the hands or for the body (for example day creams, night creams, creams or sun oils, body milks), a makeup composition (for example foundation) or a

artificial tanning composition.

  When the composition of the invention is an emulsion, the proportion of fatty substances which it contains can range from 5% to 80% by weight, and preferably from 5% to% by weight relative to the total weight of the composition. The fatty substances and the emulsifiers used in the composition in the form of an emulsion are chosen from those conventionally used in the cosmetic or pharmaceutical field. As fatty substances which can be used in the invention, there may be mentioned mineral oils (petrolatum), vegetable oils (liquid fraction of shea butter) and their hydrogenated derivatives, animal oils, synthetic oils (perhydrosqualene), silicone oils (dirmethylpolysiloxane) and fluorinated oils. As other fatty substances, mention may also be made of fatty alcohols (alcohol

  cetyl, stearyl alcohol), fatty acids (stearic acid) and waxes.

  The emulsifiers can be present in the composition in a proportion ranging from 0.3% to 30% by weight, and preferably from 0.5 to 30% by weight, per

  relative to the total weight of the composition.

  In known manner, the cosmetic or pharmaceutical compositions of the invention may also contain adjuvants customary in the corresponding fields, such as hydrophilic or lipophilic gelling agents, preservatives, antioxidants, solvents, perfumes, fillers, filters and coloring matters. Furthermore, these compositions can contain hydrophilic or lipophilic active agents. The amounts of these various adjuvants or active ingredients are those conventionally used in the cosmetic or pharmaceutical field, and for example from 0.01% to 20% of the total weight of the composition. These adjuvants or these active agents, depending on their nature, can be introduced into the fatty phase, into the aqueous phase and / or into lipid vesicles. Among the active agents which the compositions of the invention may contain, mention may in particular be made of active agents having an effect on the treatment of wrinkles or fine lines, and in particular keratolytic active agents. By keratolytic is meant an active agent having desquamating, exfoliating or exfoliating properties, or an active

  able to soften the stratum corneum.

  Among the active agents having an effect on the treatment of wrinkles or fine lines that the compositions of the invention may contain, mention may in particular be made of:

hydroxy acids and retinoids.

  The hydroxy acids can be for example co-hydroxy acids or f-

  hydroxy acids, which can be linear, branched or cyclic, saturated or unsaturated. The hydrogen atoms of the carbon chain can, in addition, be substituted by halogens, halogenated, alkylated, acylated radicals,

  acyloxylated, carbonylated or alkoxylated alkoxy having from 2 to 18 carbon atoms.

  The hydroxy acids which can be used are in particular glycolic, lactic, malic, tartaric, citric, 2-hydroxy alkanoic, mandelic, salicylic acids, as well as their alkylated derivatives such as n-octanoyl-5-salicylic acid,

  N-dodecanoyl-5-salicylic acid, n-decanoyl-5-salicylic acid, n-octyl- acid

  -salicylic acid, n-heptyloxy-5 or -4-salicylic acid, 2-hydroxy3-methyl-

  benzoic acid, or their alkoxylated derivatives such as 2-hydroxy-3- acid

  methoxybenzoic. The retinoids can in particular be retinoic acid (all-trans or 13-cis) and its derivatives, retinol (vitamin A) and its esters such as retinol palmitate,

  retinol acetate and retinol propionate, and their salts.

  These assets can be used in particular at concentrations ranging from

  0.0001% to 5% by weight relative to the total weight of the composition.

  When the compositions of the invention are intended for oral application, they can be in the usual dosage forms in this field, such as tablets, capsules, drinkable products, especially consisting of extemporaneously, granules, powders , in the usual excipients for such an application. When the compositions of the invention are intended for injection, they may be in the form of solutions containing the excipients usually used for injections, and for example in the form of a

  isotonic sodium chloride solution.

  The following examples and compositions illustrate the invention without limiting it in any way. In the compositions the proportions indicated are

  percentages by weight, unless otherwise stated.

  Example 1: Preparation of an Iris Pallida Extract: Undifferentiated iris pallida cells cultured in vitro under axenic conditions are recovered after culturing in an Erlenmeyer flask or in a fermenter by filtration through a 50 μm sieve. To 55 g of fresh material thus obtained, 27.5 ml of demineralized water are added. The whole is ground in a Turax at 24,000 rpm for 1 minute at 4 ° C. (ice bath). The ground material is centrifuged 15 to 10,000 G at 4 C. The supernatant

  is filtered at 0.22 pm (sterilizing filtration).

  The extract thus prepared is stored at 4 C. It contains approximately 15 g of material

dry per liter.

  If the plant material is from the whole plant, the fresh material to be treated is brought back according to the dry weight in order to put itself in the same extraction conditions as for in vitro. The different parts of the plant are removed according to the relative weight of each part of it. The cold treatment makes it possible to freeze the enzymatic activities, the sterilizing filtration avoids the degradation of the active ingredients by the microorganisms of the environment. Finally, the water vehicle is compatible with ex vivo receptors and facilitates formulations

cosmetic or pharmaceutical.

  Example 2: Measurement of the Affinity of the Iris Pallida Extract for the Glycine and GABA-A Receptors (Yaminobutiric Acid Type A Receptor): The affinity for the glycine receptors was determined according to the method described by Marvizon et al in Mol. Pharmacol., 1986, 30: 590-597. The affinity for GABA-A receptors was determined according to the method

  described by Snodgrass, S.R., Nature, 1978, 273: 392.

  Summary of the experimental conditions: Glycine receptor: The displacement of tritiated strychnine fixed on the glycine receptor is measured by prior incubation of 2 nM of tritiated strychnine with membranes of

  rat spinal cord for 10 minutes at 0 C.

  The iris pallida extract is tested at 1% and 5% of its initial concentration.

  The IC50 of strychnine (dose necessary to displace 50% of the tritiated strychnine attached to the receptor) is measured by displacement with unmarked strychnine. GABA-A receptor: The displacement of tritiated muscimol fixed on the GABA-A receptor is measured by prior incubation of 2.5 nM of tritiated muscimol with cerebral cortex.

rat for 10 minutes at 4 C.

  The iris pallida extract is tested at 1% and 5% of its initial concentration.

  The IC50 of muscimol (dose required to move 50% of the tritiated muscimol attached to the

  receptor) is measured by displacement with unmarked muscimol.

  Results: expressed as a percentage of inhibition of binding of the radioactive ligand Iris pallida extract (concentration) 1% 5% Glycine receptor 71 92 GABA-A receptor 100 100 Reference: strychnine (glycine) IC50o: 26.9 nM Reference: muscimol (GABA-A) IC50: 8.2 nM These results show that the iris pallida extract is a good receptor ligand

with glycine and GABA-A.

  The relationship between these receptors, chlorine channels and contractions

  makes iris pallida extract a good anti-wrinkle agent.

  EXAMPLE 3 Measurement of the Effect of the Iris Pallida Extract on the Time Period for the Appearance of an Antagonistic Effect of the Chlorine Channels (Convulsions) Induced by Strychnine in

  mouse after subcutaneous administration.

  The measurement is carried out according to the method described by Krall et al.,

Epilepsia, 1978, 19, 409-428.

  The iris pallida extract is administered subcutaneously in a volume of ml / kg, at the desired doses. 30 minutes after treatment, a solution of

  strychnine is injected subcutaneously at a dose of 1 mg / kg.

  The onset of convulsions is observed for 30 minutes after the injection of the

strychnine.

  The time taken for convulsions to appear is also measured.

  The reference product is Diazepam 3 mg / kg.

  Results: The time to onset of convulsions is expressed in seconds. Values

  data are averages of the times measured by animal in each group.

The vehicle is sterile water.

  Product Time Variation (%) Vehicle 266 Diazepam 3431 + 1190 Iris pallida (1 ml / kg) 333 + 25 Iris pallida (5 ml / kg) 321 + 21 The iris pallida extract present, delaying the time of appearance convulsions,

  a relaxing power which makes it an anti-wrinkle active.

  Example 4: examples of compositions according to the invention.

  Composition 1: Face care lotion Extract from Example 1 7.00% Antioxidant 0.05% Preservative 0.30% Ethanol (solvent) 8.00% Water qs 100% The lotion obtained acts on wrinkles during '' repeated use (application

twice a day for a month).

  Composition 2: Gel for facial care Extract from Example 1 7.00% Hydroxypropylcellulose * 1.00% Preservative 0.30% Ethanol (solvent) 15.00% Antioxidant 0.05% Water qs 100%

  *: Klucel H sold by the company Hercules (gelling agent).

  The gel obtained acts on wrinkles. It can be applied daily morning and evening

for a month.

  Composition 3: Face care cream (oil-in-water emulsion) Extract from Example 1 5.00% Glycerol stearate (emulsifier) 2.00% Polysorbate 60 (Tween 60 sold by the company ICI) (emulsifier) 1.00% Stearic acid 1.40% Triethanolamine (neutralizer) 0.70% Carbomer (Carbopol 940 sold by the company Goodrich) 0.40% Liquid fraction of shea butter 12.00% Perhydrosqualene 12.00% Preservative 0, 30% Perfume 0.50% Antioxidant 0.05% Water qs 100% We obtain a white, creamy cream, which acts on wrinkles and fine lines, and

  that can be applied daily.

  Composition 4: Face care cream (oil-in-water emulsion) Extract from Example 1 5.00% Mono-, glycerol distearate 2.00% Cetyl alcohol 1.50%

  Cetylstearyl alcohol / acetylstearyl alcohol mixture

  ethylene 33 EO 7.00% Dimethylpolysiloxane 1.50% Vaseline oil 17.50% Preservative 0.30% Perfume 0.50% Glycerin 12.50% Water qs 100%

Claims (12)

  1. Use in a cosmetic composition or for the preparation of a pharmaceutical composition, of an effective amount of an extract of at least one Iridaceae to relax and / or relax the skin and / or subcutaneous tissue. 2. Use according to claim 1, characterized in that an effective amount of an extract of at least one Iridaceae is used to relax and / or relax the   skin and / or subcutaneous tissue to reduce wrinkles and / or fine lines.   3. Use according to any one of the preceding claims, characterized   in that the cosmetic and / or pharmaceutical composition is packaged in a form suitable for administration by topical route, by oral route and / or by injection.   4. Use according to any one of the preceding claims,   characterized in that the extract is an extract of the whole plant.   5. Use according to any one of claims 1 to 3, characterized in that   that the extract is an extract of plant material obtained by in vitro culture.   6. Use according to claim 5, characterized in that the plant material   is obtained by in vitro culture of undifferentiated cells.   7. Use according to any one of the preceding claims, characterized   in that said extract is an extract of plant material from Iridaceae of a genus chosen from Romulea, Crocus, Iris, Gladiolus, Sisyrinchium or Hermodactylus. 8. Use according to claim 7, characterized in that said extract is a   extract of plant material from iris.   9. Use according to claim 8, characterized in that said extract is a extract from Iris pallida.   10. Use according to any one of the preceding claims,   characterized in that in the cosmetic composition, said Irridaceous extract is used in an amount representing from 0.01% to 20% of the total weight of the   composition and preferably from 0.1% to 5% of the total weight of the composition.   11. Use according to any one of claims 1 to 9, characterized in that   that in the pharmaceutical composition, said locust extract is used in an amount representing from 0.01% to 30% of the total weight of the composition and preferably in an amount representing from 0.1% to 10% of the total weight of the composition.   12. Method for the cosmetic treatment of wrinkles and / or fine lines, consisting in applying topically, injecting or ingesting a composition comprising   an effective amount of at least one extract of an Iridaceae.